ABSTRACT Typing of human papillomaviruses (HPV) by DNA hybridization procedures, such as reverse line blot (RLB) assay, is sensitive and well validated. However, the application these assays to high-throughput analyses limited. Here, we describe development multiplex papillomavirus genotyping (MPG), a quantitative procedure for identification multiple high- low-risk genital HPV genotypes in single reaction. MPG based on amplification general primer PCR (GP5+/6+) subsequent detection products...

Human papillomavirus (HPV) DNA detection and typing are important for diagnosis management of HPV-associated diseases. One the most commonly used PCR methods, GP5+/6+, shows weaknesses in amplifying certain types. To circumvent this limitation, we developed validated broad-spectrum primers targeting GP5+/6+ region. The addition eight upstream two downstream BSGP5+/6+ (BS) improved amplification plasmids 14 genital HPV types 10- to 1,000-fold versus without altering sensitivity 10 others. For...

Background Anal intraepithelial neoplasia (AIN), a human papillomavirus (HPV)-associated potential precursor lesion of anal cancer, is frequent among immunodeficiency virus (HIV)-positive men who have sex with (MSM). There paucity data published on the progression high-grade AIN to invasive cancer as well clinical and virological characteristics comparing margin canal carcinoma. Objectives To search for carcinoma in large series HIV-positive MSM, assess treatment response carcinoma, analyse...

Use of false cell lines remains a major problem in biological research. Short tandem repeat (STR) profiling represents the gold standard technique for line authentication. However, mismatch repair (MMR)-deficient are characterized by microsatellite instability, which could force allelic drifts combination with selective outgrowth otherwise persisting side lines, and, thus, likely to be misclassified STR profiling. On basis high-throughput Luminex platform, we developed 24-plex single...

PCR methods enable the detection of a large variety human papillomavirus (HPV) genotypes that infect anogenital tract. However, with consensus primers, general and, to lesser extent, broad-spectrum primers may underrepresent true prevalence HPV, especially multiple infections. We compared rate HPV positivity determined by BSGP5+ and BSGP6+ (BS-PCR) coupled an established bead-based multiplex genotyping (MPG) assay type-specific (TS-PCR) newly developed MPG for 735 selected cervical scraping...

Oropharyngeal squamous cell carcinomas (OPSCC) that are associated with human papilloma virus (HPV) infection carry a more favorable prognosis than those HPV-negative. However, it remains unclear which biomarker(s) can reliably determine OPSCC specimens truly driven by HPV infection. In this study, we analyzed 199 fresh-frozen for DNA, viral load, RNA expression patterns typical cervical (CxCaRNA(+)), and the HPV-targeted tumor suppressor protein p16(INK4a) as markers set of specimens, there...

The purpose of this study was to assess the clinical and radiographic long-term outcomes patients treated with a third-generation cemented total shoulder replacement followed for at least ten years.The results thirty-nine arthroplasties were analyzed clinically use Constant score on radiographs in two projections, special regard glenoid component loosening, mean eleven years (range, fifteen years) postoperatively.The 27 points 11 54 points) preoperatively 61 21 86 postoperatively (p <...

Unnoticed cell culture contamination by viruses, Mycoplasma, or other lines is not uncommon and a threat to laboratory safety the quality of scientific results. We developed validated novel high-throughput Multiplex Contamination Test (McCT), which currently able detect 37 markers in single reaction. The assay based on multiplex PCR with target-specific primers subsequent hybridization amplimers specific oligonucleotide probes. McCT proved be highly specific, sensitive robust, allows analyze...

Abstract Eight HPV types ( HPV26 , 53, 66, 67, 68, 70, 73 and 82) that are phylogenetically closely related to 12 WHO ‐defined high‐risk HR ) have been rarely but consistently identified as single infections in about 3% of cervical cancer CxCa tissues. Due lack biological data, these referred probable/possible (p) HR‐HPV . To analyse their activity direct comparison types, we selected 55 formalin‐fixed, paraffin‐embedded FFPE tissues harbouring pHR‐HPV (2–13 cases per type) 266 (7–40 from a...

Judging the carcinogenicity of human papillomavirus (HPV) types rarely found in cervical cancer (CxCa) is hindered by lack studies their biological activity tissues. To asses transcriptional HPV types, we have developed ultra-short amplimer, splice-site specific, E6*I mRNA RT-PCR assays for 12 high-risk (HR)-HPV (IARC Group 1) and eight probable/possible (pHR)-HPV 2A/B carcinogens). Previously unreported splice sites six pHR-HPV 26, 53, 67, 70, 73 82 were identified cloning sequencing. We...

Multiple human papillomavirus (HPV) genotypes often coexist within cervical epithelia and are frequently detected together in smears of different grades neoplasia. Describing the association between multiple infections disease is important generating hypotheses regarding its pathogenesis. We analyzed prevalence HPV their attribution to a screening population 999 consecutive BD SurePath liquid-based cytology samples enriched with atypical squamous cells undetermined significance (ASCUS) (n =...

Human papillomavirus (HPV) is a causal factor in virtually all cervical and subset of oropharyngeal squamous cell carcinoma (OP-SCC), whereas its role laryngeal (L-SCC) unclear. Formalin-fixed paraffin-embedded (N=154) deep-frozen tissues (N=55) 102 L-SCC patients were analysed for the presence 51 mucosal HPV types. DNA-positive (HPV DNA+) cases E6*I mRNA transcripts high risk (HR)/probably/possibly (p)HR-HPV identified, type 16 (HPV16) viral load. Expression p16INK4a, pRb, cyclin D1 p53 was...

Inverted papillomas are rare tumors of the lower urinary tract. Among 1829 reclassified bladder, renal pelves, ureters, and urethra, there were 40 (2.2%) inverted papillomas. The great majority these localized in bladder. Because distinctive histologic features patterns growth it is possible to differentiate between two basic types which termed "trabecular" "glandular." trabecular type consists widely branched, anastomosing cords urothelial cells originating directly from overlying...

Abstract High‐risk human papillomavirus (hrHPV) infection is the major risk factor for cervical cancer (CxCa). The role of genetic susceptibility in disease has been suggested, but existing data lack consistency. We conducted a nested case‐control study on 973 CxCa cases and 1,763 matched controls, from two Swedish population‐based cohorts to examine association common variants with risk. Human leukocyte antigen (HLA) alleles 24 other polymorphisms 14 genes were selected basis reported or...

Infections with high-risk human papillomaviruses (HPV), mainly HPV type 16, can cause malignant transformation of the cervical epithelium and development cancer (CxCa). A rapid precise diagnosis precancerous lesions by conventional cytology or DNA tests remains difficult often leads to overtreatment. We quantitatively analyzed HPV16 transcriptome 80 DNA-positive scrapes classified as mild cytologic grade, including no intraepithelial lesion malignancy (NIL/M; normal, n=25) low-grade squamous...

Viral oncogene RNA expression is regarded as reliable biomarker to identify oropharyngeal squamous cell carcinomas (OPSCC) with active HPV16 involvement. This study addressed whether the profile of cellular proteins p16INK4a, pRb, Cyclin D1 and p53 provide surrogate marker combinations that OPSCC in situations where only formalin-fixed biopsies are available. Protein was analyzed by immunohistochemistry on tissue microarrays created from 188 which DNA status had been established previously...

Abstract Of the 120 known human papillomaviruses (HPV), 51 HPV types infect genital mucosa. Very little is about prevalence and viral load of majority these low‐risk (Lr‐) in screening populations. We determined three subtypes 999 consecutive BD‐SurePath™ liquid‐based cervical cytology samples collected during routine gynecological health checks from Belgian women. This series was enriched with ASC‐US ( n = 100), low‐grade squamous intraepithelial lesion LSIL 100) high‐grade (HSIL) 97)...

Infections with high-risk human papillomaviruses (Hr-HPV) can cause malignant transformation of the cervical epithelium. HPV DNA tests generally are very sensitive to detect neoplastic lesions but also identify transient infections. As a consequence, specificity and positive predictive value low.We analyzed viral load Hr- possibly Hr-HPV types more than seven orders magnitude (on log10 scale) in 999 consecutive BD-SurePath liquid-based cytology samples from routine screening enriched...

To investigate the frequency of Human papillomavirus (HPV)-driven head and neck squamous cell carcinoma (HNSCC) among patients living in North-East Italy, by assessing HPV-DNA positivity all tumors additional markers whenever possible. HPV types, viral load, RNA, HPV16/18 E6 protein p16INK4a pRb expression were determined primary tumor tissues from 247 HNSCC patients. Tumor-specific seropositivity was analyzed 102 Tumor prevalence 8.5% overall (21/247) 27% oropharynx (17/63). HPV16 accounted...